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Biomedical subjects

Y K Kim

Publications and source records attributed to Y K Kim.

At least 109 records · Page 6Linked to original sources

Requirement for either a host- or pectin-induced pectate lyase for infection of Pisum sativum by Nectria hematococca.

Fungal pathogens usually have multiple genes that encode extracellular hydrolytic enzymes that may degrade the physical barriers in their hosts during the invasion process. Nectria hematococca, a plant pathogen, has two inducible pectate lyase (PL) genes (pel) encoding PL that can help degrade the carbohydrate barrier in the host. pelA is induced by pectin, whereas pelD is induced only in planta. We show that the disruption of either the pelA or pelD genes alone causes no detectable decrease in virulence. Disruption of both pelA and pelD drastically reduces virulence. Complementation of the double disruptant with pelD gene, or supplementation of the infection droplets of the double disruptant with either purified enzyme, PLA, or PLD, caused a recovery in virulence. These results show that PL is a virulence factor. Thus, we demonstrate that disruption of all functionally redundant genes is required to demonstrate the role of host barrier-degrading enzymes in pathogenesis and that dismissal of the role of such enzymes based on the effects of single-gene disruption may be premature.

Ascomycota↗

Role of lipid peroxidation and poly(ADP-ribose) polymerase activation in oxidant-induced membrane transport dysfunction in opossum kidney cells.

This study was undertaken to examine the role of lipid peroxidation and poly(ADP-ribose) polymerase (PARP) activation in H(2)O(2)-induced inhibition of Na(+)-dependent phosphate (Na(+)-Pi) uptake in opossum kidney (OK) cells. H(2)O(2) inhibited Na(+)-Pi uptake in a dose-dependent manner. H(2)O(2)-induced inhibition of Na(+)-Pi uptake was prevented by dithiothreitol and glutathione. A potent antioxidant, DPPD, had no effect on H(2)O(2) inhibition of Na(+)-Pi uptake, despite completely inhibiting lipid peroxidation induced by H(2)O(2). However, in primary cultured rabbit proximal tubular cells, the effect of H(2)O(2) on Na(+)-Pi uptake was significantly prevented by DPPD, suggesting a species difference in the role of lipid peroxidation in the inhibition of Na(+)-Pi uptake occurring with H(2)O(2). t-Butylhydroperoxide (tBHP) caused the inhibition of Na(+)-Pi uptake that was prevented by DPPD in OK cells and rabbit proximal tubular cells. The PARP inhibitor 3-aminobenzamide completely protected the inhibition of Na(+)-Pi uptake induced by H(2)O(2) but not by tBHP. H(2)O(2)-induced ATP depletion was prevented by 3-aminobenzamide but not by DPPD. tBHP-induced ATP depletion was prevented by DPPD, whereas it was not altered by 3-aminobenzamide. Effects of H(2)O(2) and tBHP on Na(+)-Pi uptake and ATP depletion were prevented by an iron chelator, deferoxamine, suggesting that the oxidants inhibit Na(+)-Pi uptake through an iron-dependent mechanism. The extent of DNA damage by tBHP was similar to that by H(2)O(2). These results indicate that the effect of H(2)O(2) on membrane transport function in OK cells is associated with PARP activation but not lipid peroxidation, whereas the effect of tBHP is associated with lipid peroxidation.

Animals↗

Role of peroxiredoxins in regulating intracellular hydrogen peroxide and hydrogen peroxide-induced apoptosis in thyroid cells.

Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. One mechanism for this action involves modulation of hydrogen peroxide (H(2)O(2))-mediated cellular responses. This report examines the expression of Prx I and Prx II in thyroid cells and their roles in eliminating H(2)O(2) produced in response to thyrotropin (TSH). Prx I and Prx II are constitutively expressed in FRTL-5 thyroid cells. Prx I expression, but not Prx II expression, is stimulated by exposure to TSH and H(2)O(2). In addition, methimazole induces a high level of Prx I mRNA and protein in these cells. Overexpression of Prx I and Prx II enhances the elimination of H(2)O(2) produced by TSH in FRTL-5 cells. Treatment with 500 micrometer H(2)O(2) causes apoptosis in FRTL-5 cells as evidenced by standard assays of apoptosis (i.e. terminal deoxynucleotidyl transferase deoxyuridine triphosphate-biotin nick end labeling, BAX expression, and poly(ADP-ribose) polymerase cleavage. Overexpression of Prx I and Prx II reduces the amount of H(2)O(2)-induced apoptosis measured by these assays. These results suggest that Prx I and Prx II are involved in the removal of H(2)O(2) in thyroid cells and can protect these cells from undergoing apoptosis. These proteins are likely to be involved in the normal physiological response to TSH-induced production of H(2)O(2) in thyroid cells.

Animals↗

Protein-protein interaction among hnRNPs shuttling between nucleus and cytoplasm.

Heterogeneous nuclear ribonucleoproteins (hnRNPs) are involved in several RNA-related biological processes such as transcription, pre-mRNA processing, mature mRNA transport to the cytoplasm, and translation. About 20 major hnRNPs from A1 to U are known. Among them, hnRNP A, D, E, I, and K are known to shuttle between the nucleus and the cytoplasm. hnRNP E2 has been seen to stabilize alpha-globin mRNA and to enhance polioviral mRNA translation. hnRNP K modulates transcription and translation of some mRNAs. hnRNP I and its homologue hnRNP L have been suggested to enhance translation of some IRES-dependent mRNAs. In order to better understand the molecular mechanisms of the biological functions of hnRNPs, we investigated protein-protein interactions of six hnRNPs (hnRNP A1, C1, E2, I, K, and L) using the yeast two-hybrid system and in vitro co-precipitation assays. All of the hnRNPs tested exerted homomeric interactions, and hnRNP E2, I, K, and L interacted with each other. In the case of hnRNP E2 and hnRNP K, the N-terminal half of the proteins containing two KH (K homologous) domains were required for protein-protein interaction, and the second quarter of hnRNP I and hnRNP L containing RRM2 (RNA recognition motif 2) was essential for protein-protein interaction. hnRNP A1 and C1 did not form complexes with other hnRNPs in our assay systems. This suggests that the hnRNPs could fall into two groups: one group, including hnRNP A1 and C1, involved in hnRNP core complex formation and another group, including hnRNP E2, I, K, and L, involved in a variety of RNA-related biological processes. Different combinations of the proteins of the second group may facilitate different biological processes in conjunction with other factors.

Amino Acid Motifs↗

Cross-linking of CD4 induces cytoskeletal association of CD4 and p56lck.

A membrane glycoprotein CD4 functions as a co-receptor of a T lymphocyte. The co-receptor function has been attributed to a protein tyrosine kinase, p56lck, which is activated upon CD4 binding to MHC molecule. In this study, we present evidences that one of the pathways through which CD4 transmits its signal is cytoskeleton association of p56lck tyrosine kinase as well as CD4 itself. Cytoskeletal association of both proteins is inhibited by a tyrosine kinase inhibitor, genistein, indicating that tyrosine protein kinase activation is important for cytoskeletal association of CD4 and p56lck. Cytoskeletal association of these proteins by CD4 cross-linking is not affected by inhibitors of protein kinase C nor PI3-kinase. Taken together, these results suggest that CD4 cross-linking activates a tyrosine kinase which then induces the simultaneous association of CD4 and p56lck with cytoskeleton.

CD4 Antigens↗

Antifungal activity of magnolol and honokiol.

Two neolignan compounds, magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl, 1) and honokiol (5,5'-diallyl-2,4'-dihydroxybiphenyl, 2), were isolated from the stem bark of Magnolia obovata and evaluated for antifungal activity against various human pathogenic fungi. Compound 1 and 2 showed significant inhibitory activities against Trichophyton mentagrophytes, Microsporium gypseum, Epidermophyton floccosum, Aspergillus niger, Cryptococcus neoformans, and Candida albicans with minimum inhibitory concentrations (MIC) in a range of 25-100 microg/ml. Therefore, compound 1 and 2 could be used as lead compounds for the development of novel antifungal agents.

Antifungal Agents↗

Intracellular pH in isolated rat renal papillary thin limbs of Henle's loop.

Intracellular pH (pHi) was measured in isolated, nonperfused and perfused rat papillary thin limbs of Henle's loops in N-2-hydroxyethylpiperazine-N'-2-ethansulfonic acid (HEPES)- or HEPES/bicarbonate-buffered medium at pH 7.4 using the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). Resting pHi was about 6.7 in descending thin limbs (DTL) and about 6.9 in ascending thin limbs (ATL), even with a medium pH of 7.4. These values appeared to reflect the acid pH of the blood in the neighboring vasa recta found in vivo. The resting pHi did not differ whether or not the medium contained bicarbonate although the total buffering capacity of the tubule cells was increased in the presence of bicarbonate. In nonperfused DTL and ATL, pHi was further acidified following an NH4Cl pulse. The rate of recovery of pHi from this level to the resting pHi was reduced by Na+ removal from the bath in both DTL and ATL and by the addition of ethylisopropylamiloride (EIPA) to the bath in the presence of Na+ in DTL. The rate of recovery was not affected by Cl- removal from the bath or K+ (75 mM) or 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS) addition to the bath in either DTL or ATL. These results suggest that the common, amiloride-sensitive, basolateral Na+/H+ exchanger plays a role in the regulation of pHi in rat papillary DTL but that a different basolateral Na+/H+ exchanger or a luminal Na+/H+ exchanger is important in rat papillary ATL.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Exercise therapy effect on natural killer cell cytotoxic activity in stomach cancer patients after curative surgery.

OBJECTIVE: To evaluate the effect of early exercise therapy on the natural killer cell cytotoxic activity (NKCA) of patients who had undergone curative resection of stomach cancer. DESIGN: Prospective study. PATIENTS: Thirty-five stomach cancer patients who had undergone curative surgery were randomly divided into an exercise group (n = 17) and a control group (n = 18). INTERVENTION: From postoperative day 2, moderated exercise using arm and bicycle ergometers performed twice a day, 5 times a week, for 14 days. The intensity of exercise was 60% of maximal heart rate. Venous blood samples were obtained on postoperative days 1, 7, and 14. MAIN OUTCOME MEASURE: Mean sequential change of NKCA. RESULTS: The mean sequential change of NKCA decreased until postoperative day 7 and then increased. Mean NKCA of day 7 decreased in both groups, compared with that at postoperative day 1. At day 14, the mean NKCA of the exercise group demonstrated a significant increase compared with that of the control group (p < .05). CONCLUSION: This study suggests that early moderate exercise has a beneficial effect on the function of in vitro NK cells in stomach cancer patients after curative surgery.

Adult↗

Intraoral removal of the submandibular gland: a new surgical approach.

The transcervical procedure used for treating sialolithiasis in the submandibular gland has been generally accepted by head and neck surgeons. However, several clinical problems after surgery through the transcervical approach have been described. We introduce a new surgical approach for excision of the submandibular gland indicated in the chronically inflamed salivary gland with or without calculus and benign mixed tumor of the submandibular gland. Thirty-one cases of submandibular gland excision through the intraoral approach were reviewed, analyzing surgical technique and morbidity. Early postoperative discomforts developed, such as a temporary lack of function of lingual nerve and a temporary limitation of tongue movement, but recovery was within a short period of time in all patients involved. No symptomatic late complications appeared, such as residual inflammation of Wharton's duct and neurologic sequelae. We suggest that this approach can be extended to the excision of the submandibular gland as an alternative to the transcervical approach. The major advantages of this approach are the avoidance of an external scar and injury to the marginal mandibular nerve or the hypoglossal nerve.

Adolescent↗

Activation of caspase cascades in Korean mistletoe (Viscum album var. coloratum) lectin-II-induced apoptosis of human myeloleukemic U937 cells.

Mistletoe lectins are of high biological activity and exert cytotoxic effects. We have previously shown that Korean mistletoe, Viscum album var. coloratum, lectin-II specifically induces apoptotic cell death in cancer cells, not normal lymphocytes. The destructive mechanism by mistletoe lectins on tumor cells was mediated by activation of c-JUN N-terminal kinase (JNK)/stress-activated protein kinase. Herein, we investigated the involvement of caspase cascade and its proteolytic cleavage effects on biosubstrates of human myeloleukemic U937 cells by D-galactoside and N-acetyl-galactosamine-specific Korean mistletoe lectin-II. Mistletoe lectin-II induced ladder pattern DNA fragmentation and activation of caspase-3, -8, and -9 of U937 cells, but not caspase-1 protease, in a time- and dose-dependent manner. Consistent with catalytic activation of protease, both poly(ADP-ribose) polymerase (PARP) and protein kinase C-delta (PKC-delta) are also cleaved in mistletoe lectin-II-treated U937 cells. An inhibitor of caspase-3-like protease, DEVD-CHO peptide, significantly inhibited mistletoe lectin-II-induced apoptosis, PARP cleavage, and fragmentation of DNA. These results provide the evidence that Korean mistletoe lectin-II induces apoptotic death of U937 cells via activation of caspase cascades.

Adjuvants, Immunologic↗

Hippocampal lesions enhance startle gating-disruptive effects of apomorphine in rats: a parametric assessment.

Prepulse inhibition of the startle reflex is an operational measure of sensorimotor gating that is impaired in schizophrenia patients and dopamine agonist-treated rats. Previous reports demonstrated an enhanced sensitivity to the prepulse inhibition-disruptive effects of the D(1)/D(2) agonist apomorphine in adult rats four weeks after cytotoxic lesions of the hippocampus, but left unanswered several important questions regarding the nature of this apparent lesion-induced dopamine supersensitivity. Because of the potential importance of this model to current theories of the pathophysiology of schizophrenia, studies now assessed specific features of this effect of hippocampus lesions on prepulse inhibition in rats. The enhanced prepulse inhibition-disruptive effects of apomorphine in ventral hippocampus-lesioned rats were unaffected by startle pulse intensity, suggesting an independence of this lesion effect from potential ceiling effects of elevated startle magnitude. These lesion effects were observed four weeks post-lesion, but not two weeks post-lesion, suggesting a delayed development of this phenomenon. No enhancement of apomorphine sensitivity was observed in rats four weeks after lesions restricted to the dorsal hippocampus; in contrast, these lesions significantly increased "no-drug" levels of prepulse inhibition. Ventral hippocampus-lesioned rats exhibited a significant reduction in prepulse inhibition after subthreshold doses of either the selective D(2)-family agonist quinpirole or the partial D(1) agonist SKF 38393, suggesting that activation of either receptor family is adequate for the expression of this effect of ventral hippocampus lesions. This may be an important paradigm for understanding the contribution of ventral hippocampus dysfunction to the neurobiology of impaired sensorimotor gating in neuropsychiatric populations.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

Inhibitor of tumor necrosis factor-alpha production in lipopolysaccharide-stimulated RAW264.7 cells from Amorpha fruticosa.

Certain flavonoids were reported to show an immunoregulatory activity against lymphocyte proliferation and cytokine production. In the course of a search for tumor necrosis factor (TNF)-alpha inhibitory compounds from natural plants, we also isolated a prenylfavanone type of flavonoid, amoradicin, from the extract of Amorpha fruticosa by activity-guided fractionation. This compound significantly inhibited TNF-alpha production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells with an IC(50) value of 28.5 microM. The activity was comparable or higher than those of standard flavonoid compounds, genistein and silybin with IC(50) of 24.9 and 140.3 microM, respectively.

Animals↗

Citrus red mite (Panonychus citri) is a common sensitizing allergen among children living around citrus orchards.

BACKGROUND: A recent investigation has suggested that Panonychus citri (citrus red mite) is the most common causative allergen in citrus-cultivating farmers with asthma or allergic rhinitis. OBJECTIVE: A cross-sectional survey was performed to evaluate the prevalence of asthma and sensitization to P. citri among children living around citrus orchards. METHODS: A total of 1,055 children living around citrus orchards were enrolled. There were 544 boys and 511 girls with a mean age of 12.3 (range 7 to 16) years. They were evaluated by the ISAAC questionnaire, and a skin prick test with 13 common aeroallergens including P. citri and Japanese cedar pollen. Children who recorded ever having experienced wheezing on the questionnaire were evaluated by methacholine bronchial provocation test and determined to have asthma if they showed bronchial hyperresponsiveness to methacholine simultaneously. RESULTS: The positive skin test response to one or more inhalant allergens except P. citri was 35.5%, but when including P. citri, the positive response increased to 37.7%. The most common sensitizing allergens in the order of decreasing frequency were Dermatophagoides pteronyssinus (26.6%), D. farinae (22.7%), P. citri (14.2%), cockroach (11.3%), and Japanese cedar pollen (9.7%). Among the children without sensitization to house dust mites, the sensitization rate to P. citri was significantly higher in those 13 and over than in those under 12 (5.8% versus 2.5%, P < .05). The prevalence of asthma was not different between the children with sensitization to P. citri and those without it (5.4% versus 4.0%, P > .05), although the prevalence was higher in those with sensitization to D. pteronyssinus or D. farinae than in those without sensitization to these allergens (7.9% versus 2.9%, P < .05; 9.7% versus 2.6%, P < .05). CONCLUSION: Citrus red mite is a common sensitizing allergen among children living around citrus orchards. The features of sensitization and development of asthma are different between citrus red mite and house dust mites.

Administration, Inhalation↗

Two-spotted spider mite (Tetranychus urticae): an important allergen in asthmatic non-farmers symtomatic in summer and fall months.

BACKGROUND: The Tetranychus urticae (two-spotted spider mite, TSM) is a spider mite commonly found on fruit trees, herbaceous plants, and greenhouse flowers. OBJECTIVE: To evaluate the clinical significance of TSM-derived allergens in non-farmers with asthma living around pear orchards, and to assess the allergenic relationship with house dust mite. METHODS: Skin prick tests with TSM were performed in 50 asthmatic non-farmers living around pear orchards. The serum TSM-specific IgE was measured in the 16 asthmatics with a positive skin response to TSM. To diagnose TSM-induced asthma, specific bronchial challenges with TSM were performed in the 16 asthmatics. ELISA inhibition tests were performed to assess the allergenic cross-reactivity with house dust mites. RESULTS: Ten of the 16 asthmatics with positive skin responses to TSM showed a significant bronchoconstriction following inhalation of TSM. Eight of the ten TSM-induced asthmatics had high serum TSM-specific IgE and one showed a positive skin response only to TSM. Their asthmatic symptoms were aggravated seasonally, especially in summer and early fall. On ELISA inhibition tests, partial inhibitions with D. pteronyssinus antigens were noted in sera from five asthmatics with positive skin responses to both TSM and D. pteronyssinus, although no inhibition was noted in serum from an asthmatic with a positive skin response only to TSM. CONCLUSION: Tetranychus urticae may be an important allergen in asthmatic non-farmers living around pear orchards and TSM extracts contain species-specific allergens as well as commonly shared allergens with house dust mite.

Adolescent↗

Sensitivity to citrus red mite and the development of asthma.

BACKGROUND AND OBJECTIVE: The citrus red mite (Panonychus citri, CRM) is an important allergen in the development of asthma and/or rhinitis in citrus farmers and a common sensitizing allergen among exposed children. The aim of this study was to evaluate CRM-induced skin responses and serum-specific IgE, IgG4, and IgG1 responses, and their role in the development of asthmatic symptoms among exposed adolescents. METHODS: A total of 100 adolescents between the ages of 15 and 16 years (59 males and 41 females) were randomly recruited. All of the subjects responded to the ISAAC questionnaire themselves. Skin prick tests to CRM were carried out and serum-specific IgE, IgG4, and IgG1 levels were measured using ELISA. RESULTS: Fourteen subjects (14%) had experienced wheezing during last 12 months. The wheal size formed by CRM was larger than 3 mm in 23 subjects (23%). Serum-specific IgE levels were elevated in 21 subjects (21%), IgG4 levels were elevated in 32 subjects (32%), and IgG1 levels were elevated in five subjects (5%). The prevalence of wheezing during last 12 months was significantly higher in subjects with positive skin responses and serum-specific IgE responses induced by CRM than in those with negative skin responses and low serum IgE responses (30.4% versus 9.0%, P < .05; 33.3% versus 8.7%, P < .05). The prevalence of wheezing, however, did not differ according to serum-specific IgG4 levels (9.4% versus 16.2%, P > .05). Skin responses to CRM showed a significant correlation with serum-specific IgE levels, but not with IgG1 and IgG4 levels. Serum CRM-specific IgE levels correlated with specific IgG1 and IgG4 levels, but no significant correlation was found between serum IgG1 and IgG4 levels. CONCLUSION: The specific IgE responses induced by CRM were prevalent among adolescents living near citrus farms, and may be important in the development of asthma. The specific IgG4 and IgG responses induced by CRM play a minimal role in the development of asthma symptoms.

Adolescent↗

Efficiency of promoter and cell line in high-level expression of erythropoietin.

Efficiency of viral promoters and various cell lines in directing high-level expression of human erythropoietin (Epo) was investigated. To investigate the effects of various viral promoters and cell lines on the Epo expression level, genomic Epo with the 5' and 3' untranslated regions (UTRs) deleted was cloned next to the simian virus 40 early promoter, cytomegalovirus early promoter or SRalpha promoter. These expression vectors were transfected into COS-7, BHK-21 and Chinese hamster ovary (CHO)/dhfr(-) cells, respectively. The COS-7 cells transfected with the vector containing the SRalpha promoter showed the highest expression level ( approximately 103 IU/ml) at 72 h post-transfection. For the development of Epo-producing stable cell lines, BHK-21 and CHO/dhfr(-) cells transfected with the 5',3'-UTR-deleted genomic Epo under the control of the SRalpha promoter were cultured with media containing zeocin. Several clones of zeocin-resistant BHK-21 and CHO/dhfr(-) cells were cultured in the presence of methotrexate (MTX). A BHK-21 clone selected in the presence of 500 nM MTX expressed and secreted approximately 490 IU/ml Epo into the medium. A CHO/dhfr(-) clone selected in the presence of 20 nM MTX expressed and secreted approximately 45 IU/ml Epo into the medium. Southern-blot analysis indicated that enhancement of Epo expression in the MTX-resistant stable cells might be related to the amplification of gene copy number.

Animals↗

Bronchial responsiveness to methacholine is increased in citrus red mite (Panonychus citri)-sensitive children without asthmatic symptoms.

BACKGROUND: A recent investigation has suggested that citrus red mite (Panonychus citri, CRM) is a common sensitizing allergen among children living around citrus farms. OBJECTIVE: A cross-sectional survey was performed to evaluate the bronchial responsiveness to methacholine in CRM-sensitive children without asthmatic symptoms. SUBJECTS AND METHODS: A total of 375 children living around citrus farms were enrolled in this study. There were 201 boys and 174 girls with a mean age of 12.5 (range 7-16) years. Each subject was evaluated by a questionnaire, a skin prick test with 13 common inhalant allergens including CRM, and a methacholine bronchial provocation test (MBPT). RESULTS: Positive rate of MBPT (PC20-methacholine < 25 mg/mL) was higher in atopic rhinitic and atopic control children than in nonatopic rhinitic and nonatopic control subjects (41.3% and 33.3% vs 17.2% and 15.4%, respectively, both P < 0.05). Slope of dose-response curve (DRS, %/mg/mL) was higher in atopic rhinitic children than in nonatopic rhinitic, atopic control and nonatopic control subjects (14.3 +/- 0.87 vs 9.4 +/- 0.72, 10.0 +/- 1.37 and 9.2 +/- 1.51, P < 0.05, respectively). Both the positive rate of MBPT and the DRS were increased in children with positive skin response to CRM than in those without sensitization (48.2% vs 22.9%, P = 0.0001; 15.6 +/- 1.26 vs 10.2 +/- 0.65, P = 0.0001). CONCLUSION: Bronchial responsiveness is increased in citrus red mite-sensitive children even if they have no asthmatic symptoms.

Adolescent↗